Tyrosine 537 within the Na+,K+-ATPase α-subunit is essential for AP-2 binding and clathrin-dependent endocytosis

Stefania Cotta Doné, Ingo B. Leibiger, Riad Efendiev, Adrian I. Katz, Barbara Leibiger, Per Olof Berggren, Carlos H. Pedemonte, Alejandro M. Bertorello

Research output: Contribution to journalArticle

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Abstract

In renal epithelial cells endocytosis of Na+,K+-ATPase molecules is initiated by phosphorylation of its α1-sub-unit, leading to activation of phosphoinositide 3-kinase and adaptor protein-2 (AP-2)/clathrin recruitment. The present study was performed to establish the identity of the AP-2 recognition domain(s) within the Na+,K+-ATPase α1-subunit. We identified a conserved sequence (Y537LEL) within the α1-subunit that represents an AP-2 binding site. Binding of AP-2 to the Na+,K+-ATPase α1-subunit in response to dopamine (DA) was increased in OK cells stably expressing the wild type rodent α-sub-unit (OK-WT), but not in cells expressing the Y537A mutant (OK-Y537A). DA treatment was associated with increased α1-subunit abundance in clathrin vesicles from OK-WT but not from OK-Y537A cells. In addition, this mutation also impaired the ability of DA to inhibit Na+,K+-ATPase activity. Because phorbol esters increase Na+,K+-ATPase activity in OK cells, and this effect was not affected by the Y537A mutation, the present results suggest that the identified motif is specifically required for DA-induced AP-2 binding and Na+,K+-ATPase endocytosis.

Original languageEnglish
Pages (from-to)17108-17111
Number of pages4
JournalJournal of Biological Chemistry
Volume277
Issue number19
DOIs
StatePublished - May 10 2002
Externally publishedYes

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Clathrin
Endocytosis
Protein Binding
Tyrosine
Adenosine Triphosphatases
Dopamine
Proteins
Adaptor Protein Complex Subunits
Mutation
Phosphorylation
1-Phosphatidylinositol 4-Kinase
Conserved Sequence
Phorbol Esters
Phosphatidylinositols
sodium-translocating ATPase
Rodentia
Carrier Proteins
Phosphotransferases
Epithelial Cells
Chemical activation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Doné, S. C., Leibiger, I. B., Efendiev, R., Katz, A. I., Leibiger, B., Berggren, P. O., ... Bertorello, A. M. (2002). Tyrosine 537 within the Na+,K+-ATPase α-subunit is essential for AP-2 binding and clathrin-dependent endocytosis. Journal of Biological Chemistry, 277(19), 17108-17111. https://doi.org/10.1074/jbc.M201326200

Tyrosine 537 within the Na+,K+-ATPase α-subunit is essential for AP-2 binding and clathrin-dependent endocytosis. / Doné, Stefania Cotta; Leibiger, Ingo B.; Efendiev, Riad; Katz, Adrian I.; Leibiger, Barbara; Berggren, Per Olof; Pedemonte, Carlos H.; Bertorello, Alejandro M.

In: Journal of Biological Chemistry, Vol. 277, No. 19, 10.05.2002, p. 17108-17111.

Research output: Contribution to journalArticle

Doné, SC, Leibiger, IB, Efendiev, R, Katz, AI, Leibiger, B, Berggren, PO, Pedemonte, CH & Bertorello, AM 2002, 'Tyrosine 537 within the Na+,K+-ATPase α-subunit is essential for AP-2 binding and clathrin-dependent endocytosis', Journal of Biological Chemistry, vol. 277, no. 19, pp. 17108-17111. https://doi.org/10.1074/jbc.M201326200
Doné, Stefania Cotta ; Leibiger, Ingo B. ; Efendiev, Riad ; Katz, Adrian I. ; Leibiger, Barbara ; Berggren, Per Olof ; Pedemonte, Carlos H. ; Bertorello, Alejandro M. / Tyrosine 537 within the Na+,K+-ATPase α-subunit is essential for AP-2 binding and clathrin-dependent endocytosis. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 19. pp. 17108-17111.
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